Team:Tsinghua/Notebook/26 July 2010


Module I, Group 2c

To test the effectivity of newly-synthetized primer, PCR assay to amplify mCherry, Kan and eGFP, Chlr is conducted.

PCR system (FastPFU):

 H2O	                       31.5μl
 5×buffer	               10μl
 dNTP	                       5μl
 primer1	               1μl
 primer2	               1μl
 template	               1μl
 FastPFU	               0.5μl
 Total	                       50μl

PCR program----

 1.95℃	                       1min
 2.95℃ 	               20s
 3.53℃for eGFP  	        
   51℃for mCherry	       20s
   51℃for Kan&Chlr	       
 4.72℃	                       30s
 30 cycles from 2 to 4
 5.72℃                       5min

Result for PCR: After agarose gel is run, no specific bands are observed.

Conclusion: reaction condition for PCR is not ideal enough, thus we shall change the condition and try this again.